# US 10,869,949 B2 · Method for treating subjects suffering from chronic ulcers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,869,949 B2 - Document ID: US10869949B2 - Status: Granted - Kind: B2 - Application number: 16/845,633 - Filed: 2020-04-10 - Published: 2020-10-15 - Granted: 2020-12-22 - Earliest claimed: 2019-04-11 - Assignees: Geistlich Pharma AG; University of Southern California - Applicants: Geistlich Pharma AG; University of Southern California - Inventors: Charles Zelen; David Armstrong; Paul Glat; Jarrod Kaufman; Marco Mehr; Lothar Schloesser; Mark Spilker - Web page: https://patentgrep.com/patent/US10869949B2 ## CPC - A61L 27/24: Collagen - A61L 27/3808: {Endothelial cells} - A61L 27/3813: {Epithelial cells, e.g. keratinocytes, urothelial cells} - A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells} - A61L 27/44: having a macromolecular matrix - A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)} - A61L 27/56: Porous materials, {e.g. foams or sponges} - A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like - A61L 2300/414: Growth factors - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract A method, material, and kit for promoting neutrophils and monocytes to localize at a chronic ulcer site, promoting formation of a multi-layered cell structure in the ulcer site, promoting conversion of monocytes to macrophages, promoting secretion of the patient's own growth factors, promoting tissue proliferation and cell migration, promoting production and cross-linking of collagen at the chronic ulcer site, promoting growth of endothelial cells, promoting angiogenesis that was stalled at the chronic ulcer site, promoting formation of a vascular network and granulation, promoting oxygenation of the chronic ulcer site, and reducing one or more of purulent drainage, erythema, pain, warming, tenderness, induration, and bleeding at the chronic ulcer site. ## Claims ### Claim 1 A method of treating a chronic skin ulcer in a subject in need thereof, comprising: i) cleaning to remove bacteria and other pathogens and/or debriding the chronic skin ulcer until the edges of the ulcer contain viable tissue; ii) aseptically implanting into the chronic skin ulcer of the subject a multilayer sheet of collagen material in dry state comprising (a) a barrier layer of collagen material having a smooth face and a rough fibrous face opposite said smooth face and (b) a spongeous matrix layer of collagen material connected to said rough fibrous face, said spongeous matrix layer of collagen material having an open sponge-like texture, such that the rough fibrous face of said barrier layer of collagen material to which is connected said spongeous matrix layer of collagen material having an open sponge-like texture, faces toward and is adjacent to the bed of the chronic skin ulcer; iii) hydrating the implanted multilayer sheet of collagen material in dry state using blood, an isotonic solution or a combination thereof; and iv) providing a dressing over the implanted, hydrated multilayer sheet of collagen material, thereby restarting stalled cell migration, proliferation and angiogenesis at the chronic skin ulcer site. ### Claim 2 (depends on 1) The method of claim 1, wherein the collagen of said barrier layer of collagen material is predominantly collagen I, collagen Ill or a mixture thereof. ### Claim 3 (depends on 1) The method of claim 1, wherein said the collagen of said spongeous matrix layer of collagen material is predominantly collagen I, collagen Ill or a mixture thereof. ### Claim 4 (depends on 1) The method of claim 1, wherein multilayer sheet of collagen material has a thickness of about 0.5-25 mm. ### Claim 5 (depends on 1) The method of claim 1, wherein the chronic ulcer extends at least through the dermis and has been present for greater than 4 weeks. ### Claim 6 (depends on 1) The method of claim 1, wherein the chronic ulcer extends at least through the hypodermis and has been present for greater than 6 weeks. ### Claim 7 (depends on 1) The method of claim 1, further comprising applying a secondary dressing or re-dressing the chronic ulcer after step iv) is performed. ### Claim 8 (depends on 1) The method of claim 1, further comprising applying sterile saline to remove a dressing material from the multilayer sheet of collagen material after step iv) is performed. ### Claim 9 (depends on 1) The method of claim 1, further comprising changing the dressing over the implanted multilayer sheet of collagen material every 1 to 7 days after step iv) is performed. ### Claim 10 (depends on 1) The method of claim 1, further comprising removing exudate from the chronic ulcer site every 1 to 7 days after step iv) is performed. ### Claim 11 (depends on 1) The method of claim 1, further comprising inspecting the chronic ulcer every 1 to 7 days after step iv) and removing the dressing after a first visible epithelialization is observed at the chronic ulcer or removing the implanted multilayer sheet of collagen material and repeating steps i) to iv) if one or more of redness, swelling, hematomas, blistering, inflammation, excess exudate, infection, and necrosis are observed at the chronic ulcer. ### Claim 12 (depends on 1) The method of claim 1, further comprising performing one or more of toe-blood pressure readings, pulse volume recordings, transcutaneous oxygen measurements, and skin perfusion pressure measurements. ### Claim 13 (depends on 1) The method of claim 1, further comprising one or more of promoting neutrophils and monocytes to localize at the chronic ulcer site, promoting formation of a multi-layered cell structure in the ulcer site, promoting conversion of monocytes to macrophages, promoting secretion of the patient's own growth factors, promoting tissue proliferation and cell migration, promoting production and cross-linking of collagen at the chronic ulcer site, promoting growth of endothelial cells, promoting angiogenesis that was stalled at the chronic ulcer site, promoting formation of a vascular network and granulation, promoting oxygenation of the chronic ulcer site, and reducing one or more of purulent drainage, erythema, pain, warming, tenderness, induration, and bleeding at the chronic ulcer site. ### Claim 14 A method of increasing liquid uptake capacity in a chronic skin ulcer of a subject in need thereof, comprising: i) aseptically implanting into the chronic skin ulcer of the subject a multilayer sheet of collagen material in dry state comprising (a) a barrier layer of collagen material having a smooth face and a rough fibrous face opposite said smooth face and (b) a spongeous matrix layer of collagen material connected to said rough fibrous face, said spongeous matrix layer of collagen material having an open sponge-like texture, such that said rough fibrous face of said barrier layer of collagen material to which is connected said spongeous matrix layer of collagen material having an open sponge-like texture faces toward and is adjacent to the bed of the chronic skin ulcer; and ii) hydrating the multilayer sheet of collagen material in dry state using blood, an isotonic solution or a combination thereof, thereby increasing liquid uptake capacity in the chronic skin ulcer. ### Claim 15 (depends on 14) The method of claim 14, further comprising inhibiting exudate drainage, bleeding from the chronic ulcer, and preventing floating away of the multilayer sheet of collagen material out of the bed of the chronic ulcer. ### Claim 16 A method of promoting hemostasis in a chronic skin ulcer of a subject in need thereof, comprising: i) aseptically implanting into the chronic skin ulcer of the subject a multilayer sheet of collagen material in dry state comprising (a) a barrier layer of collagen material having a smooth face and a rough fibrous face opposite said smooth face and (b) a spongeous matrix layer of collagen material connected to said rough fibrous face, said spongeous matrix layer of collagen material having an open sponge-like texture, such that said rough fibrous face of said barrier layer of collagen material to which is connected said spongeous matrix layer of collagen material having an open sponge-like texture faces toward and is adjacent to the bed of the chronic skin ulcer; and ii) hydrating the implanted multilayer sheet of collagen material using blood, an isotonic solution or a combination thereof, thereby promoting hemostasis in the chronic skin ulcer. ### Claim 17 (depends on 16) The method of claim 16, wherein blood clot formation in a chronic ulcer is accelerated by at least 2-fold compared to blood clot formation in a chronic ulcer in the absence of said implanted multilayer sheet of collagen material. ### Claim 18 A method of binding and preserving a subject's growth own factors in a chronic skin ulcer of a subject in need thereof, comprising: i) aseptically implanting into the chronic skin ulcer of the subject a multilayer sheet of collagen material in dry state comprising (a) a barrier layer of collagen material having a smooth face and a rough fibrous face opposite said smooth face and (b) a spongeous matrix layer of collagen material connected to said rough fibrous face, said spongeous matrix layer of collagen material having an open sponge-like texture, such that said rough fibrous face of said barrier layer of collagen material to which is connected said spongeous matrix layer of collagen material having an open sponge-like texture faces toward and is adjacent to the bed of the chronic ulcer; and ii) hydrating the implanted multilayer sheet of collagen material using blood, an isotonic solution or a combination thereof, thereby promoting binding of said subject's own growth factors with the multilayer sheet of collagen material and preservation of said subject's own growth factors and growth factor activity in the chronic skin ulcer thereby inducing expression of one or more growth factor-responsive genes in one or more human cell types in the chronic skin ulcer of the subject. ### Claim 19 (depends on 18) The method of claim 18, wherein the growth factors are two or more of transforming growth factors (TGFs), fibroblast growth factors (FGFs), epidermal growth factor (EGF), Insulin-like Growth Factor (IGF-1), Platelet-derived Growth Factors (PDGFs), and vascular endothelial growth factors (VEGFs). ### Claim 20 (depends on 18) The method of claim 18, wherein said one or more human cell types are human fibroblasts, human epidermal keratinocytes, human endothelial cells and human pluripotent stem cells. ### Claim 21 (depends on 1) The method of claim 1, further comprising attracting one or more human cell types to the chronic skin ulcer, wherein said one or more human cell types are human fibroblasts, human epidermal keratinocytes, human endothelial cells and human pluripotent stem cells. ### Claim 22 (depends on 1) The method of claim 1, further comprising promoting attachment and growth of one or more human cell types in the chronic skin ulcer, wherein said one or more human cell types are human fibroblasts, human epidermal keratinocytes, human endothelial cells and human pluripotent stem cells. ### Claim 23 (depends on 1) The method of claim 1, further comprising inhibiting one or more MMPs in the chronic skin ulcer, wherein the MMPs are a plurality of MMP-1, MMP-2, MMP-3, MMP-8, and MMP-9. ### Claim 24 (depends on 1) The method of claim 1, wherein the subject suffers from diabetic foot ulcer (DFU) or venous leg ulcer (VLU). ### Claim 25 (depends on 1) The method of claim 1, wherein the subject has been or is being treated with corticosteroid therapy, is undergoing radiation therapy, is receiving anti-coagulation therapy, chemotherapy, or uses drugs, alcohol, tobacco, or other agents that disrupt a normal ulcer healing process. ### Claim 26 (depends on 1) The method of claim 1, further comprising treating the subject with compression therapy, vacuum assisted closure (VAC), offloading, negative pressure, hyperbaric oxygen therapy, or a combination thereof. ### Claim 27 (depends on 1) The method of claim 1, wherein the multilayer sheet of collagen material in dry state has physical properties such that it absorbs about 7 to about 12 times its weight of biological fluids. ### Claim 28 (depends on 1) The method of claim 1, wherein the multilayer sheet of collagen material has not been artificially cross-linked, has not had any growth factors or other ulcer-treating agents added to it, and/or has not had any antimicrobial agents added to it. ### Claim 29 (depends on 1) The method of claim 1, further comprising, after 4 to 7 days, removing at least a portion of the implanted multilayer sheet of collagen material and repeating the method steps. ### Claim 30 (depends on 1) The method of claim 1, further comprising providing a pH of or about 3.5 to about 6.5 in the chronic ulcer site.